It is possible that by the following method, and induced to differentiate easily induced pluripotent stem cells or embryonic stem cells in large quantities. By using the substrate surface having a cell non-adhesive area around the cell attachment regions, Allowed to attach to the cell attachment regions of the substrate surface by inducing differentiation of medium under (1) the cell, (2) Then, grown to confluence in cell attachment on the surface of the cells, The further continued (3) culture, is stratified into three-dimensional from the boundary between the non-adherent cell area of cell adhesion area surrounding the cells, and, Bound stratification plurality of portions formed on the cell adhesion region and (4) finally, to produce a cell mass with thick cell adhesion region.次の方法により、大量の胚性幹細胞或いは人工多能性幹細胞を簡便に分化誘導することができる。細胞付着性領域の周囲に細胞非付着性領域を有する基材表面を利用し、(1)当該細胞を分化誘導化培地下で基材表面の細胞付着性領域に付着させ、(2)その後、当該細胞を細胞付着性表面上でコンフルエントになるまで増殖させ、(3)培養をさらに継続し、当該細胞を細胞付着性領域周囲の細胞非付着性領域との境界部から3次元に重層化させ、及び、(4)最終的にその細胞付着性領域内に形成された複数の重層化部分を結合させ、細胞付着性領域全体を厚みのある細胞集合体を作製する。
